Motor Neuron Diseases: Cellular and Animal Models

Haase, Georg

doi:10.1002/3527600906.mcb.200400156

Cited by 2 publications

(2 citation statements)

References 134 publications

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“…Furthermore, the reduction and replacement of animal use in research should be a target for all researchers. An alternative is cell culture, but most in vitro models to study the neuromuscular system are monocultures of skeletal muscle cells (SkMCs), frequently of animal origin [ 10 , 11 ], and the absence of functional innervation and lack of NMJ formation in these models result in an incomplete replication of in vivo conditions [ 12 ]. Given the importance of innervation and functional NMJs for muscle differentiation and function [ 13 ], models where NMJs are generated may considerably enhance understanding of neuromuscular pathologies and sarcopenia and provide a platform for the validation of new treatments.…”

Section: Introductionmentioning

confidence: 99%

A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction

Saini

Faroni

Reid

et al. 2020

JCM

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Background: In many neurodegenerative and muscular disorders, and loss of innervation in sarcopenia, improper reinnervation of muscle and dysfunction of the motor unit (MU) are key pathogenic features. In vivo studies of MUs are constrained due to difficulties isolating and extracting functional MUs, so there is a need for a simplified and reproducible system of engineered in vitro MUs. Objective: to develop and characterise a functional MU model in vitro, permitting the analysis of MU development and function. Methods: an immortalised human myoblast cell line was co-cultured with rat embryo spinal cord explants in a serum-free/growth fact media. MUs developed and the morphology of their components (neuromuscular junction (NMJ), myotubes and motor neurons) were characterised using immunocytochemistry, phase contrast and confocal microscopy. The function of the MU was evaluated through live observations and videography of spontaneous myotube contractions after challenge with cholinergic antagonists and glutamatergic agonists. Results: blocking acetylcholine receptors with α-bungarotoxin resulted in complete, cessation of myotube contractions, which was reversible with tubocurarine. Furthermore, myotube activity was significantly higher with the application of L-glutamic acid. All these observations indicate the formed MU are functional. Conclusion: a functional nerve-muscle co-culture model was established that has potential for drug screening and pathophysiological studies of neuromuscular interactions.

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Section: Introductionmentioning

confidence: 99%

A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction

Saini

Faroni

Reid

et al. 2020

JCM

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“…For instance, most studies on NM diseases rely on in vivo animal models that do not entirely reflect disease in humans 1. Shortcomings of in vitro models of NM disorders are that they are largely based on cells derived from animals,2,3 or skeletal muscle cell (SkMC) culture systems that fail to mimic in vivo conditions, particularly due to the lack of functional innervation 4. Thus, the development of new models to study and manipulate NMJs has the potential to provide significant insight into NM disease pathogenesis and detection, and to test the efficacy of innovative therapies.…”

Section: Introductionmentioning

confidence: 99%

<p>Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells</p>

Saini¹,

Faroni²,

Samid³

et al. 2019

SCCAA

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Background Neuromuscular junctions (NMJs) consist of the presynaptic cholinergic motoneuron terminals and the corresponding postsynaptic motor endplates on skeletal muscle fibers. At the NMJ the action potential of the neuron leads, via release of acetylcholine, to muscle membrane depolarization that in turn is translated into muscle contraction and physical movement. Despite the fact that substantial NMJ research has been performed, the potential of in vivo NMJ investigations is inadequate and difficult to employ. A simple and reproducible in vitro NMJ model may provide a robust means to study the impact of neurotrophic factors, growth factors, and hormones on NMJ formation, structure, and function. Methods This report characterizes a novel in vitro NMJ model utilizing immortalized human skeletal muscle stem cells seeded on 35 mm glass-bottom dishes, cocultured and innervated with spinal cord explants from rat embryos at ED 13.5. The cocultures were fixed and stained on day 14 for analysis and assessment of NMJ formation and development. Results This unique serum- and trophic factor-free system permits the growth of cholinergic motoneurons, the formation of mature NMJs, and the development of highly differentiated contractile myotubes, which exhibit appropriate configuration of transversal triads, representative of in vivo conditions. Conclusion This coculture system provides a tool to study vital features of NMJ formation, regulation, maintenance, and repair, as well as a model platform to explore neuromuscular diseases and disorders affecting NMJs.

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Motor Neuron Diseases: Cellular and Animal Models

Cited by 2 publications

References 134 publications

A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction

A Novel Bioengineered Functional Motor Unit Platform to Study Neuromuscular Interaction

<p>Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells</p>

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